This research is part of an effort to better understand the molecular mechanisms underlying human nervous system development and function, as well as the pathogenesis of certain neurogenetic disorders. Our studies have focused on structural and active site properties of the human non-neuronal and neuron specific enolases, lysosomal hydrolases (glucocerebro- sidase and Alpha-galactosidase A), other enzymes (particularly those peptides and proteins that interact with excitable membranes), and venom toxins. Proteins are purified from both human and animal tissues using affinity chromatography, electrophoretic separation, and high performance liquid chromatography. Using microsequencing techniques, the complete amino acid sequence of glucocerebrosidase, and major portions of sequences for the neuronal and non-neuronal enolases, venom toxins, and Alpha-galactosidase A have been obtained. Peptide maps of both normal and mutant proteins are generated using chemical (cyanogen bromide) and enzymatic (trypsin, thermolysin, V8 protease) cleavage. The identification of carbohydrate attachment sites, sulfhydryl residues, and intrachain disulfide residues is used to predict protein structure. Alkylating agents and enzyme inhibitors are used to define active sties. From the primary protein sequence, hydrophobic and hydrophilic domains of the protein are identified. Information obtained from these protein structure studies permits the design of oligonucleotides and peptides that are synthesized for collaborative research involving antibody production, cDNA cloning, DNA sequence analysis and in vitro mutagenesis.